Nontoxic clostridium perfringens and clostridium septicum fusion protein vaccine and production method thereof

Abstract

The invention relates to a nontoxic clostridium perfringens and clostridium septicum fusion protein vaccine and a production method thereof. The fusion protein vaccine prepared by the method adopts aclostridium perfringens epsilon toxin which is subjected to codon optimization and contains three amino acid mutations, and spoilage which contains four amino acid mutations and eleven amino acid deletions, namely not only kept the integrity and spatial conformation of the natural toxin are to the maximum extent, but also the biological safety hazard caused by single amino acid mutation is avoided. The vaccine also has the advantages of simple preparation process, excellent vaccine efficacy and the like, greatly reduces the biological safety risk in the vaccine production process. Compared with the current commercialized sheep fast-epidemic and sheep enterotoxemia inactivated vaccines in China, the vaccine is an ideal candidate vaccine for upgrading existing clostridium perfringens and clostridium septicum vaccines in China; in addition, when combined vaccines are prepared by the vaccine together with other antigens, the combined vaccine can be prepared without increasing the use doseof the combined vaccine.

Description

technical field [0001] The invention relates to an avirulent Clostridium perfringens and Clostridium putrefaction fusion protein vaccine and a production method thereof. It belongs to the field of veterinary biological products. Background technique [0002] Clostridium perfringens and Clostridium putrefaction are anaerobic bacteria that can cause disease in humans and various animals, and are extremely harmful to human health and livestock and poultry breeding. Among them, Clostridium perfringens is one of the main pathogens of traumatic gas gangrene and human food poisoning, sudden gangrene of sheep, lamb dysentery, necrotic enteritis of cattle and sheep, and enterotoxemia of cattle and sheep. The main pathogen of the epidemic. The pathogenic factors of both Clostridia are exotoxins secreted by the bacteria. Clostridium perfringens can be divided into A, B, C, D, E according to the types of four main lethal exotoxins α (CPA), β (CPB), ε (ETX) and ι (CPI) produced. Five...

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Problems solved by technology

Although there have been reports of attenuated mutants of Clostridium perfringens epsilon toxin, such as Yu Li et al. ("Clostridium perfringens epsilon toxin attenuated mutants and their applications", application No.: 201410707351.6) mutated the tyrosine at position 71 of the mature toxin of the wild-type Clostridium perfringens epsilon toxin into a non-aromatic amino acid, and obtained the epsilon toxin mutant attenuated for cells or animals. Since this application only Mutation of 1 amino acid is prone to reverse mutation and becomes a highly virulent toxin again, which will bring serious biosafety risks to the actual large-scale production of vaccines in the future

China Animal Disease Prevention and Control Center Song Xiaohui et al. ("Recombinant ε protein for inhibiting Clostridium perfringens infection and its preparation method and application", application number: 201610301989.9) designed 3 recombinant ε toxin proteins, but all of them lacked the wild type The first 50 amino acid residues of the epsilon toxin protein, this strategy of using protein fragments as immunogens is likely to result in incomplete antigenicity of the antigenic protein, thereby affecting its immunogenicity

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